We used an improved procedure to analyze the intraflagellar transport (IFT) of protein particles in Chlamydomonas and found that the frequency of the particles, not only the velocity, changes at each end of the flagella. Thus, particles undergo structural remodeling at both flagellar locations. Therefore, we propose that the IFT consists of a cycle composed of at least four phases: phases II and IV, in which particles undergo anterograde and retrograde transport, respectively, and phases I and III, in which particles are remodeled/exchanged at the proximal and distal end of the flagellum, respectively. In support of our model, we also identified 13 distinct mutants of flagellar assembly (fla), each defective in one or two consecutive phases of the IFT cycle. The phase I-II mutant fla10-1 revealed that cytoplasmic dynein requires the function of kinesin II to participate in the cycle. Phase I and II mutants accumulate complex A, a particle component, near the basal bodies. In contrast, phase III and IV mutants accumulate complex B, a second particle component, in flagellar bulges. Thus, fla mutations affect the function of each complex at different phases of the cycle.
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2 April 2001
Article|
March 26 2001
Protein Particles in Chlamydomonas Flagella Undergo a Transport Cycle Consisting of Four Phases
Carlo Iomini,
Carlo Iomini
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
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Veronica Babaev-Khaimov,
Veronica Babaev-Khaimov
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
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Massimo Sassaroli,
Massimo Sassaroli
bDepartment of Physiology and Biophysics, Mount Sinai School of Medicine, New York, New York 10029
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Gianni Piperno
Gianni Piperno
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
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Carlo Iomini
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
Veronica Babaev-Khaimov
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
Massimo Sassaroli
bDepartment of Physiology and Biophysics, Mount Sinai School of Medicine, New York, New York 10029
Gianni Piperno
aDepartment of Cell Biology and Anatomy, Mount Sinai School of Medicine, New York, New York 10029
Abbreviations used in this paper: Ab, antibody; DHC, dynein heavy chain; IFT, intraflagellar transport; KHP, kinesin homologue protein; ts, temperature-sensitive; VE-DIC, video-enhanced differential interference contrast.
Received:
August 02 2000
Revision Requested:
January 30 2001
Accepted:
January 30 2001
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 2001 The Rockefeller University Press
2001
The Rockefeller University Press
J Cell Biol (2001) 153 (1): 13–24.
Article history
Received:
August 02 2000
Revision Requested:
January 30 2001
Accepted:
January 30 2001
Citation
Carlo Iomini, Veronica Babaev-Khaimov, Massimo Sassaroli, Gianni Piperno; Protein Particles in Chlamydomonas Flagella Undergo a Transport Cycle Consisting of Four Phases. J Cell Biol 2 April 2001; 153 (1): 13–24. doi: https://doi.org/10.1083/jcb.153.1.13
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